ObjectiveTaking the cuproptosis/oxidative stress pathway as the entry point， this study investigated the effect and mechanism of Liangyi Paste on hepatic lipid deposition in naturally aged mice fed with a high-fat diet. MethodsAfter adaptive feeding， 80 ten-week-old male C57BL/6 mice were used. Thirty of them were randomly divided into three groups （10 mice per group）： The 12-month-old control group （12MCON）， the 15-month-old control group （15MCON）， and the 15-month-old group with a high-fat diet （15MHFD）. The 12MCON and 15MCON groups were continuously fed a standard diet， while the 15MHFD group started receiving a high-fat diet at 12 months of age. Tissue samples were collected at the corresponding time points for each group. The remaining 50 mice were randomly divided into five groups （10 mice per group）： the 20-month-old control group （20MCON）， the model group， and the low-， medium-， and high-dose Liangyi Paste groups （2.91 ， 5.82 ， 11.64 g·kg^-1·d^-1， respectively）. The 20MCON group was continuously fed a standard diet， while the other groups started receiving a high-fat diet at 15 months of age. At 18 months of age， the Liangyi Paste groups were administered the corresponding doses of Liangyi Paste by gavage， while the 20MCON and model groups were given an equal volume of saline by gavage. After 8 weeks of continuous gavage （when the mice reached 20 months of age）， tissue samples were collected. Hepatic TG levels were measured using assay kits； liver histology and lipid deposition were observed via hematoxylin-eosin （HE） and oil red O staining； reactive oxygen species （ROS） were detected by enzyme-linked immunosorbent assay （ELISA）； Cu²⁺， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured by colorimetry； mRNA and protein expression of genes related to cuproptosis and oxidative stress pathways were analyzed by Real-time polymerase chain reaction（Real-time PCR） and Wes automated protein expression system. ResultsCompared with 12MCON， the 15MCON group showed significantly increased hepatic TG， Cu²⁺， ROS， and MDA levels （P<0.01）， decreased SOD （P<0.01）， hepatocyte swelling， and disordered arrangement. The mRNA and protein levels of ferredoxin 1 （FDX1）， dihydrolipoamide S-acetyltransferase （DLAT）， heat shock protein 70 （HSP70）， dihydrolipoamide dehydrogenase （DLD）， pyruvate dehydrogenase E1 subunit-β （PDHB）， nuclear factor erythroid 2-related factor 2 （Nrf2）， and peroxisome proliferator-activated receptor γ （PPARγ） were significantly elevated （P<0.05， P<0.01）. Compared with 15MCON group， the 15MHFD and 20MCON groups exhibited further increases in TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， and aggravated hepatocyte swelling and disorder. There were increased lipid droplets with mild vacuolization in the 15MHFD group， and no significant lipid deposition was observed in the 20MCON group. FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and protein levels were significantly increased （P<0.05， P<0.01）. Compared with 20MCON group， the model group demonstrated markedly elevated TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， severe hepatic steatosis， and upregulated expression of FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and proteins （P<0.05， P<0.01）. All abnormalities were significantly reversed after Liangyi Paste treatment. ConclusionLiangyi paste can ameliorate hepatic lipid deposition in naturally aged mice with a high-fat diet by modulating the cuproptosis/oxidative stress pathway.

The dysregulation of cyclin-dependent kinase 12 (CDK12), which may result from genomic alterations or modulation by upstream effectors, is implicated in cancer oncogenesis and progression. CDK12 overexpression or activation is sufficient to induce tumor initiation, recurrence, and therapeutic resistance. However, CDK12 may also exert tumor-suppressive functions in a context-dependent manner. Therefore, caution is warranted when targeting CDK12 in future clinical trials. A comprehensive elucidation of the dual roles and underlying mechanisms of CDK12 in carcinogenesis is urgently needed to advance precision oncology. This review provides an overview of the current understanding of the dysregulation and biological roles of CDK12 in cancer. Subsequently, we systematically summarize the functions and mechanisms of the oncogenic and tumor-suppressive roles of CDK12 in different contexts. Finally, we discuss the potential of CDK12 as a novel therapeutic target and its implications in clinical oncology, offering insights into future directions for innovative cancer treatment strategies.

Objective To explore the safety and feasibility of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing for the treatment of inguinal indirect hernia in children≤3 months old.Methods From January 2020 to September 2024,49 children ≤ 3 months old with indirect hernia underwent external silk thread traction-assisted two-port laparoscopic internal ring suturing.All the patients were found to complicated with redundant peritoneum around internal ring.Under transumbilical single-port laparoscopic monitoring,the 3-mm laparoscopic needle holder holing the 3-0 needle-thread nylon was inserted through the inferior edge of the internal ring,entering the peritoneum and traveling through the retroperitoneal space.The external nylon thread was advanced to drag the redundant peritoneum,allowing the suture needle cross the surface of the spermatic cord vessels,the Doom's triangle,and the vas deferens(boys)or the uterine round ligament(girls)in the retroperitoneal space,followed by circular suturing and closure of the internal ring.Results The surgery time was(13.5±2.5)min for the 35 cases of unilateral hernia and(24.5±3.5)min for the 14 cases of bilateral hernia.No intraoperative complications occurred,and all the patients were discharged within 6 h post-operation.Except for 3 children of lost to follow-up,46 children were followed up for 6-26 months(mean,9.5 months),with 19 cases followed for＞12 months.No complications such as recurrence,wound infection,hydrocele,iatrogenic undescended testicle,or testicular atrophy were observed during the follow-ups.Conclusion For children ≤ 3 months old with indirect hernia,the use of 3-mm laparoscopic needle holder combined with 3-0 needle-thread nylon in external silk thread traction-assisted two-port laparoscopic internal ring suturing enhances the flexibility and convenience of the procedure,being safety and feasibility.

The dysregulation of cyclin-dependent kinase 12(CDK12),which may result from genomic alterations or modulation by upstream effectors,is implicated in cancer oncogenesis and progression.CDK12 over-expression or activation is sufficient to induce tumor initiation,recurrence,and therapeutic resistance.However,CDK12 may also exert tumor-suppressive functions in a context-dependent manner.Therefore,caution is warranted when targeting CDK12 in future clinical trials.A comprehensive elucidation of the dual roles and underlying mechanisms of CDK12 in carcinogenesis is urgently needed to advance pre-cision oncology.This review provides an overview of the current understanding of the dysregulation and biological roles of CDK12 in cancer.Subsequently,we systematically summarize the functions and mechanisms of the oncogenic and tumor-suppressive roles of CDK12 in different contexts.Finally,we discuss the potential of CDK12 as a novel therapeutic target and its implications in clinical oncology,offering insights into future directions for innovative cancer treatment strategies.

Objective To explore the targets and mechanism of Buyang Huanwu Decoction in preventing and treating atherosclerosis through network pharmacology,molecular docking and animal experiment.Methods The main active components and related targets of Buyang Huanwu Decoction were obtained and screened through databases such as TCMSP,HERB,UniProt,PubChem,SwissADME and SwissTargetPrediction.Six disease databases such as DrugBank,GeneCards,DisGeNET,OMIM,PharmGKB and TTD were used to obtain AS related targets.R language was used to obtain common targets for the drug and disease.STRING 11.5 database was used for protein interaction network analysis,and Cytoscape 3.9.1 software was used to conduct network topology parameters and obtain core targets.GO and KEGG enrichment analysis were performed on the common targets using DAVID platform.Molecular docking of core genes with the main active components of Buyang Huanwu Decoction was conducted using AutoDock Vina 1.5.6 software.An AS mouse model was established and intervened with Buyang Huanwu Decoction.The lipid content of the mouse aortic sinus was observed using Oil Red O staining.Western blot was used to detect the expression of PI3K/AKT/p38 signaling pathway,and ELISA was used to detect the contents of TNF-α and IL-17 in mouse serum.Results A total of 104 main active components and 283 action targets of Buyang Huanwu Decoction were obtained,as well as 5 347 AS related targets and 218 common targets for drugs and disease.Buyang Huanwu Decoction may use key active components such as quercetin,kaempferol and baicalin as core targets for TP53,MAPK1,AKT1,and exert its therapeutic effect on AS through PI3K-Akt signaling pathway,TNF signaling pathway and IL-17 signaling pathway,etc.Molecular docking indicated that quercetin,luteolin,kaempferol and baicalin had good affinity with TP53,HSP90AA1,MAPK1,AKT1 and RELA targets.MAPK1 had strong binding activity with baicalin and luteolin.The experimental results showed that Buyang Huanwu Decoction could reduce the lipid content in aortic sinus of AS mice,reduce the contents of TNF-α and IL-17 in serum(P<0.01)and significantly down-regulate the expression levels of PI3K,p-AKT and p-p38 proteins(P<0.05,P<0.01).Conclusion The mechanism of Buyang Huanwu Decoction for preventing and treating AS may be related to regulating the expression of the PI3K/AKT/p38 signaling pathway,thereby reducing inflammatory response.

Objective To summarize the surgical experience of residual cavity reduction by using dermal-fascial suture in giant posterior cervical subcutaneous mass(maximum diameter≥7 cm)resection.Methods From September 2019 to June 2024,27 pediatric patients with posterior cervical subcutaneous masses with maximum diameter≥7 cm underwent mass resection.By using 4-0 polypropylene suture,the residual cavity was reduced through a novel suturing technique:sutures were passed through the dermal layer into the residual cavity,and then anchored to the cervical muscular fascia through the surgical incision.After exiting the skin at 0.5-1 cm adjacent to the entry point,a 1 cm × 1 cm gauze bolster was placed between entry and exit points,and tied and secured.A drainage tube was placed at the lowest point of the residual cavity.Results All the 27 procedures succeeded.The operation time was 70-160 min(mean,95.0±12.5 min),with dermal-fascial suturing requiring 10-20 min(mean,15 min).The drainage volume was measured 2-5 ml at postoperative day 3,with no detectable fluctuation or significant tenderness at the residual cavity and skin incision.Cervical ultrasound confirmed no obvious residual cavity or fluid accumulation,and the drainage tube was removed.The postoperative hospital stay was 4-8 d(mean,6 d).Postoperative pathological diagnosis included 16 cases of lipomas,7 cases of fibrolipomas,and 4 cases of lymphangiomas.At 10 d follow-up visit,no discomfort was reported.Upon dressing change,the cervical shape was found to be normal,with no protrusions or indentations,or incision infection.At one month after surgery,ultrasound showed no abnormalities in the subcutaneous space of the neck,and the skin incision had healed.At 6 months after surgery,no recurrence of tumor was found.Conclusion Dermal-fascial suture effectively reduces residual cavity following giant posterior cervical mass resection in children,promoting tissue regeneration and wound healing while preventing complications.

OBJECTIVE To investigate the horizontal transfer mechanisms and genetic evolutionary characteristics of virulence genes in hypervirulent Klebsiella pneumoniae(HvKP)and resistance genes in carbapenem-resistant K.pneumoniae(CRKP).METHODS The donor strain was a CRKP strain harboring both VIM and KPC resist-ance genes.The positive control was a CRKP strain containing blaIMP-4.The recipient strain was Escherichia coli J53.The horizontal transfer capability of the resistance genes was verified by plasmid conjugation assay.Fifty HvKP strains were collected from the First Affiliated Hospital of Jinan University from 2017 to 2021 and conjuga-ted with the plasmids of five CRKP strains which could mediate the transmission of drug resistance genes.The conjugants were screened by selective medium containing meropenem,and polymerase chain reaction(PCR)was used to detect the resistance and virulence genes in the conjugation-successful strains.The lethality of conjugants was verified by Galleria mellonella larval assays.RESULTS The donor CRKP strain had the ability to mediate re-sistance gene transmission.Its blaKPC gene was successfully transferred to E.coli J53,but the transference of the VIM gene failed.Five CRKP strains with the ability to mediate resistance gene transmission were screened,of which one strain carried both VIM and KPC resistance genes,and the other four strains carried the IMP-4 resist-ance gene.Only one CRKP strain with the blaIMP-4 resistance gene was successfully conjugated with HvKP,carry-ing both virulence genes(aerobactin and RmpA)and the blaIMP-4 resistance gene,with positive wire drawing test result.The Galleria mellonella larvae test showed that the conjugation-successful strain caused the death of all lar-vae within 24 hours.CONCLUSIONS In the hospital environment,CRKP and HvKP can form"superbugs"with both drug resistance and high virulence through plasmid conjugation,posing a severe challenge to clinical an-ti-infection treatment.It is necessary to strengthen the prevention and control of hospital-acquired infections.

Objective To explore the safety and feasibility of extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure in the treatment of pediatric ovarian sliding hernia.Methods From September 2019 to September 2024,57 children with ovarian sliding hernia underwent extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure.Under the guidance of single-port umbilical laparoscopic monitoring,a 2-0 suturing needle with thread was inserted through the opposite internal ring and used to suture the round ligament of the uterus at the affected side's internal ring.The needle was passed through and exited near the entry point,and the extracorporeal suture traction was used to pull the ovary(and part of the fallopian tube)back into the abdominal cavity from the inguinal canal,maintaining the suture tension to prevent the ovary and fallopian tube from re-entering the internal ring.The internal ring was fully exposed.A 2-0 suturing needle with thread was inserted at the affected side(2-3 cm from the internal ring)to perform a circular suture of the peritoneum at the internal ring.The internal ring was closed at the level of internal ring orifice.Results All the 57 cases of ovarian sliding hernia were confirmed to be unilateral,with 18 cases on the right side(including 12 cases of pure ovarian sliding hernia and 6 cases of ovarian and fallopian tube sliding hernia)and 39 cases on the left side(including 22 cases of pure ovarian sliding hernia and 17 cases of ovarian and fallopian tube sliding hernia).The average surgery time was(22.5±2.5)min.All the patients were discharged within 6 h postoperatively and no complications were noted.A total of 49 patients were followed up for6-20 months(mean 9.5 months),with 21 cases followed for more than 12 months.No complications such as incision infection,recurrence of hernia,or Nuck's cyst were observed.Ultrasonic examinations showed normal bilateral ovarian blood flow.Conclusion For pediatric ovarian sliding hernia,extracorporeal suture traction of round ligament of the uterus assisted two-port laparoscopic internal ring closure can effectively retract the ovary(and part of the fallopian tube)from the inguinal canal back into the abdominal cavity,fully exposing the normal anatomical form of the internal ring,with the characteristics of safety and feasibility.

ObjectiveTo investigate the effects of modified Ditan tang on genes related to the transcription-translation feedback loop （TTFL） of biorhythm in the rat model of non-alcoholic fatty liver disease （NAFLD） and its mechanism for prevention and treatment of NAFLD. MethodsSixty-five healthy SPF male SD rats were randomly assigned into blank （n=20）， model （n=15）， and low-， medium-， and high-dose （2.68， 5.36， and 10.72 g·kg^-1·d^-1， respectively） modified Ditan tang （n=10） groups. Other groups except the blank group were fed a high-fat diet for 12 weeks. The modified Ditan tang groups were treated with the decoction at corresponding doses by gavage， and the blank and model groups were treated with an equal volume of normal saline from the 9th week for 4 weeks. The levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） in the serum were measured by an automatic biochemical analyzer. TG and non-esterified fatty acid （NEFA） assay kits were used to measure the levels of TG and NEFA in the liver. The pathological changes in the hypothalamus and liver were observed by hematoxylin-eosin staining， and the lipid deposition in the liver was observed by oil red O staining. The levels of brain-muscle ARNT-like protein 1 （BMAL1/ARNTL） in the hypothalamus and liver were determined by immunohistochemical staining. The mRNA and protein levels of BMAL1， circadian locomotor output cycles kaput （CLOCK）， period circadian clock 2 （PER2）， and cryptochrome1 （Cry1） in the hypothalamus and liver were determined by Real-time PCR and Western blot， respectively. ResultsCompared with the blank group， the model group showed elevated levels of TG， TC， LDL-C， AST， and ALT （P<0.01） and a lowered level of HDL-C （P<0.05） in the serum， elevated levels of TG and NEFA in the liver （P<0.01）， pyknosis and deep staining of hypothalamic neuron cells， and a large number of vacuoles in the brain area. In addition， the model group showed lipid deposition in the liver， up-regulated mRNA and protein levels of CLOCK and BMAL1 （P<0.01）， and down-regulated mRNA and protein levels of Cry1 and PER2 （P<0.01） in the hypothalamus and liver. Compared with the model group， all the three modified Ditan tang groups showed lowered levels of TG， TC， LDL-C， ALT， and AST （P<0.05， P<0.01） and an elevated level of HDL-C （P<0.05） in the serum， and lowered levels of TG and NEFA （P<0.05， P<0.01） in the liver. Furthermore， the three groups showed alleviated pyknosis and deep staining of hypothalamic neuron cells， reduced lipid deposition in the liver， down-regulated mRNA and protein levels of CLOCK and BMAL1 （P<0.05， P<0.01）， and up-regulated mRNA and protein levels of Cry1 and PER2 （P<0.05， P<0.01） in the hypothalamus and liver. ConclusionModified Ditan tang can reduce lipid deposition in the liver and regulate the expression of CLOCK， BMAL1， Cry1， and PER2 in the TTFL of NAFLD rats.

ObjectiveTo explore the effect of gypenosides （GPs） on liver lipid deposition in metabolism-associated fatty liver disease （MAFLD） mice and its mechanism based on classical/non-classical ferroptosis. MethodsEight male C57BL/6 mice in a blank group and 32 male apolipoprotein E gene knockout （ApoE^-/-） mice were randomly divided into a model group， a low-dose GPs （GPs-L） group， a high-dose GPs （GPs-H） group， and a simvastatin （SV） group. Starting from the second week， mice in the blank group were given a maintenance diet， and the other four groups were fed a high-fat diet daily. After eight weeks of feeding， mice in the GPs-L and GPs-H groups were given GPs of 1.487 mg·kg^-1·d^-1 and 2.973 mg·kg^-1·d^-1， respectively， and mice in the SV group were given simvastatin of 2.275 mg·kg^-1·d^-1. Mice in the blank group and the model group were given saline of equal volume by gavage for four weeks. The content of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST） in the serum of mice in each group was detected by an automatic biochemical analyzer. The level of non-esterified fatty acid （NEFA） and TG in the mouse liver was measured by the kit. The change in liver tissue structure and lipid deposition was observed by hematoxylin-eosin （HE） and oil red O staining. The levels of coenzyme Q₁₀ （CoQ₁₀）， glutathione （GSH）， malondialdehyde （MDA）， and Fe²⁺ in serum， as well as nicotinamide adenine dinucleotide phosphate ［NAD（P）H］ in the liver were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of ferroptosis suppressor protein 1 （FSP1） in the liver of mice was observed by the immunohistochemical （IHC） method， and the expression of genes and proteins related to classical and non-classical ferroptosis pathways was analyzed by real-time polymerase chain reaction （Real-time PCR） and Wes automated protein expression analysis system. ResultsCompared with those in the blank group， the levels of TC， TG， LDL-C， ALT， and AST in serum and TG and NEFA in the liver in the model group were significantly increased， and the level of HDL-C in serum was significantly decreased （P<0.01）. The liver tissue structure changed， and there were fat vacuoles of different sizes and a large number of red lipid droplets， with obvious lipid deposition. The level of CoQ10 and GSH in serum and NADH in the liver were significantly decreased， while the level of MDA and Fe²⁺ in serum was significantly increased （P<0.01）. The mRNA and protein expressions of cystine/glutamate transporter （xCT/SLC7A11）， glutathione peroxidase （GPX4）， p62， nuclear factor E₂-related factor 2 （Nrf2）， and FSP1 were significantly decreased， and the mRNA and protein expressions of tumor antigen （p53）， spermidine/spermine N1-acetyltransferase 1 （SAT1）， arachidonate 15-lipoxygenase （ALOX15）， and Kelch-like epichlorohydrin-associated protein-1 （Keap1） were significantly increased （P<0.01）. Compared with those in the model group， the level of TC， TG， LDL-C， ALT， and AST in serum and TG and NEFA in the liver of mice in the GPs-L， GPs-H， and SV groups were decreased， while the level of HDL-C in serum was significantly increased （P<0.05， P<0.01）. The liver tissue structure and lipid deposition were improved. The levels of CoQ10 and GSH in serum and NADH in the liver were significantly increased， while the levels of MDA and Fe²⁺ in serum were significantly decreased （P<0.05， P<0.01）. The mRNA and protein expressions of xCT， GPX4， p62， Nrf2， and FSP1 were significantly increased， while the mRNA and protein expressions of p53， SAT1， ALOX15， and Keap1 were significantly decreased （P<0.05， P<0.01）. ConclusionGPs can interfere with liver lipid deposition in MAFLD mice through classical/non-classical ferroptosis pathways.